Abstract
Although non-viral vectors have been explored for nearly 30 years as a potential means of treating both acquired and inherited diseases, several obstacles have limited their success as a viable treatment option. We have found that by sequentially injecting the cationic lipid and plasmid DNA in vivo, it is possible to overcome several of these obstacles. By altering the order in which the lipid and DNA interact with both each other and the blood (i.e. serum proteins), we were able to improve transgene activity and decrease the inflammatory response to the vector. In this study, we further explore the mechanism by which sequential injection is able to increase the efficacy of these lipid-based vectors. Using 2-D gel electrophoresis, we examined proteins bound to the sequentially formed complexes and compared those to proteins bound to pre-formed lipoplex. Several unique proteins have been identified and are currently under investigation for their ability to minimize the inflammatory response as well as increase the activity. This data indicates that there is potential for the development of more efficient lipid-based vectors, which contain one or more of these naturally occurring serum proteins. Supported by NIH grant AI48851.
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